Heteroaryl containing bile acid analogs as fxr/tgr5 agonists and methods of use thereof

ABSTRACT

The present invention provides compounds of Formula I, 
     
       
         
         
             
             
         
       
     
     pharmaceutical compositions comprising these compounds and methods of using these compounds to treat or prevent a disease or disorder mediated by FXR and/or TGR5.

RELATED APPLICATION

This application claims the benefit of U.S. Provisional Application No. 62/298,695, filed on Feb. 23, 2016. The entire teachings of the above application are incorporated herein by reference.

TECHNICAL FIELD

The present invention relates generally to compounds and pharmaceutical compositions useful as FXR/TGR5 modulators. Specifically, the present invention relates to bile acid derivatives and methods for their preparation and use.

BACKGROUND OF THE INVENTION

Farnesoid X Receptor (FXR) is an orphan nuclear receptor initially identified from a rat liver cDNA library (B M. Forman, et al., Cell, 1995, 81(5), 687-693) that is most closely related to the insect ecdysone receptor. FXR is a member of the nuclear receptor family of ligand-activated transcription factors that includes receptors for the steroid, retinoid, and thyroid hormones (D J. Mangelsdorf, et al., Cell, 1995, 83(6), 841-850). The relevant physiological ligands of FXR are bile acids (D. Parks et al., Science, 1999, 284(5418), 1362-1365). The most potent one is chenodeoxycholic acid (CDCA), which regulates the expression of several genes that participate in bile acid homeostasis. Farnesol and derivatives, together called farnesoids, are originally described to activate the rat orthologue at high concentration but they do not activate the human or mouse receptor. FXR is expressed in the liver, throughout the entire gastrointestinal tract including the esophagus, stomach, duodenum, small intestine, colon, ovary, adrenal gland and kidney. Beyond controlling intracellular gene expression, FXR seems to be also involved in paracrine and endocrine signaling by upregulating the expression of the cytokine Fibroblast Growth Factor (J. Holt et al., Genes Dev., 2003, 17(13), 1581-1591; T. Inagaki et al., Cell Metab., 2005, 2(4), 217-225).

Small molecule compounds which act as FXR modulators have been disclosed in the following publications: WO 2000/037077, WO 2003/015771, WO 2004/048349, WO 2007/076260, WO 2007/092751, WO 2007/140174, WO 2007/140183, WO 2008/051942, WO 2008/157270, WO 2009/005998, WO 2009/012125, WO 2008/025539, and WO 2008/025540. Further small molecule FXR modulators have been recently reviewed (R. C. Buijsman et al. Curr. Med. Chem. 2005, 12, 1017-1075).

TGR5 receptor is a G-protein-coupled receptor that has been identified as a cell-surface receptor that is responsive to bile acids (BAs). The primary structure of TGR5 and its responsiveness to bile acids has been found to be highly conserved in TGR5 among human, bovine, rabbit, rat, and mouse, and thus suggests that TGR5 has important physiological functions. TGR5 has been found to be widely distributed in not only lymphoid tissues but also in other tissues. High levels of TGR5 mRNA have been detected in placenta, spleen, and monocytes/macrophages. Bile acids have been shown to induce internalization of the TGR5 fusion protein from the cell membrane to the cytoplasm (Kawamata et al., J. Bio. Chem., 2003, 278, 9435). TGR5 has been found to be identical to hGPCR19 reported by Takeda et al., FEBS Lett. 2002,520, 97-101.

TGR5 is associated with the intracellular accumulation of cAMP, which is widely expressed in diverse cell types. While the activation of this membrane receptor in macrophages decreases pro-inflammatory cytokine production, (Kawamata, Y., et al., J. Biol. Chem. 2003, 278, 9435-9440) the stimulation of TGR5 by BAs in adipocytes and myocytes enhances energy expenditure (Watanabe, M., et al. Nature. 2006, 439, 484-489). This latter effect involves the cAMP-dependent induction of type 2 iodothyronine deiodinase (D2), which by, locally converting T4 into T3, gives rise to increased thyroid hormone activity. Consistent with the role of TGR5 in the control of energy metabolism, female TGR5 knock-out mice show a significant fat accumulation with body weight gain when challenged with a high fat diet, indicating that the lack of TGR5 decreases energy expenditure and elicits obesity (Maruyama, T., et al., J. Endocrinol. 2006, 191, 197-205). In addition, and in line with the involvement of TGR5 in energy homeostasis, bile acid activation of the membrane receptor has also been reported to promote the production of glucagon-like peptide 1 (GLP-1) in murine enteroendocrine cell lines (Katsuma, S., Biochem. Biophys. Res. Commun., 2005, 329, 386-390). On the basis of all the above observations, TGR5 is an attractive target for the treatment of disease e.g., obesity, diabetes and metabolic syndrome.

In addition to the use of TGR5 agonists for the treatment and prevention of metabolic diseases, compounds that modulate TGR5 modulators are also useful for the treatment of other diseases e.g., central nervous diseases as well as inflammatory diseases (WO 01/77325 and WO 02/84286). Modulators of TGR5 also provide methods of regulating bile acid and cholesterol homeostasis, fatty acid absorption, and protein and carbohydrate digestion.

There is a need for the development of FXR and/or TGR5 modulators for the treatment and prevention of disease.

SUMMARY OF THE INVENTION

In one aspect, the invention provides compounds represented by Formula I, and pharmaceutically acceptable salts, stereoisomers, solvates, hydrates and combinations thereof:

wherein: Ring A is a five membered heteroaromatic ring which has three heteroatoms independently selected from nitrogen, oxygen and sulfur, and can be further substituted when possible; preferably Ring A has (a) one oxygen or sulfur atom and two nitrogen atoms or (b) three nitrogen atoms; R₁ is selected from the group consisting of:

1) Hydrogen;

2) Substituted or unsubstituted —C₁-C₈ alkyl;

3) Substituted or unsubstituted —C₂-C₈ alkenyl;

4) Substituted or unsubstituted —C₂-C₈ alkynyl;

5) Substituted or unsubstituted aryl; and

6) Substituted or unsubstituted arylalkyl;

preferably R₁ is hydrogen or methyl;

m is selected from 0, 1, 2 and 3; preferably m is 0, 1 or 2; R₂ is hydrogen, hydroxyl, —OSO₃H, —OSO₃ ⁻, —OAc, —OPO₃H₂ or —OPO₃ ²⁻; preferably R₂ is hydrogen or hydroxyl; R₃ is hydrogen, halogen, CN, N₃, hydroxyl, —OSO₃H, —OSO₃ ⁻, —OAc, —OPO₃H₂, —OPO₃ ²⁻, —SR₁ or —NHR₂, wherein, R₁ is as defined previously; preferably R₃ is hydrogen; alternatively, R₂ and R₃ are taken together with the carbon atoms to which they are attached to form —CH═CH— or a cycloalkyl ring or heterocycloalkyl ring such as, but not limited to cyclopropyl or epoxide; R₄ and R₅ are each independently selected from hydrogen or hydroxyl protecting group such as, but not limited to, acetyl, trimethyl silyl, or benzyl; preferably R₄ and R₅ are both hydrogen; R₆ is selected from the group consisting of:

1) Hydrogen;

2) Halogen;

3) Substituted or unsubstituted —C₁-C₈ alkyl;

4) Substituted or unsubstituted —C₂-C₈ alkenyl;

5) Substituted or unsubstituted —C₂-C₈ alkynyl; and

6) Substituted or unsubstituted —C₃-C₈ cycloalkyl; preferably R₆ is C₁-C₄-alkyl, more preferably R₆ is ethyl;

R₇ is selected from the group consisting of:

1) Hydrogen;

2) Substituted or unsubstituted —C₁-C₈ alkyl;

3) Substituted or unsubstituted —C₂-C₈ alkenyl;

4) Substituted or unsubstituted —C₂-C₈ alkynyl;

5) Substituted or unsubstituted —C₃-C₈ cycloalkyl;

6) Substituted or unsubstituted —C₃-C₈ heterocycloalkyl;

7) Substituted or unsubstituted aryl;

8) Substituted or unsubstituted heteroaryl;

9) Substituted or unsubstituted aryl alkyl;

10) —NR₈R_(9;)

11) —N(R₁₀)C(O)R₁₁;

12) —N(R₁₀)C(O)OR₁₁;

13) —N(R₁₀)C(O)NR₈R_(9;)

14) —N(R₁₀)SO₂NR₈R_(9;)

15) —N(R₁₀)SO₂R₁₁;

16) —N(R₁₀)C(O)N(R₈)SO₂R₁₁; and

17) —N(R₁₀)C(O)N(R₈)SO₂NR₈R₉,

R₈, R_(9,) R₁₀, and R₁₁ are each independently selected from hydrogen, substituted or unsubstituted —C₁-C₈ alkyl, substituted or unsubstituted —C₂-C₈ alkenyl, substituted or unsubstituted —C₂-C₈ alkynyl, substituted or unsubstituted —C₃-C₈ cycloalkyl, substituted or unsubstituted —C₃-C₈ heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted benzyl; alternatively, R₈ and R₉ and/or R₈ and R₁₀ are taken together with the atoms to which they are attached to form an optionally substituted heteroaryl ring.

Each preferred group stated above can be taken in combination with one, any or all other preferred groups.

DETAILED DESCRIPTION OF THE INVENTION

In one embodiment of the invention is a compound represented by Formula (I) as described above, or a pharmaceutically acceptable salt, hydrate, solvate, ester or prodrug thereof.

In preferred embodiments, the compounds of the invention have the stereochemistry set forth in Formula IA:

where m, ring A, R₁, R₂, R₃, R₄, R₅, R₆, and R₇ have the meanings given for these variables in Formula I.

In one embodiment of the invention is a compound represented by Formula (I) as described above, or a pharmaceutically acceptable salt, hydrate, solvate, ester or prodrug thereof. A-R₇ can be represented by, but is not limited to, groups (I-1) to (I-11):

wherein: R_(a) is selected from the group consisting of:

1) Hydrogen;

2) Substituted or unsubstituted —C₁-C₈ alkyl;

3) Substituted or unsubstituted —C₂-C₈ alkenyl;

4) Substituted or unsubstituted —C₂-C₈ alkynyl;

5) Substituted or unsubstituted —C₃-C₈ cycloalkyl;

6) —C(O)R₁₁;

7) —C(O)OR₁₁;

8) —C(O)NR₈R₉;

9) —SO₂R₁₁; and

10) —C(O)N(R₈)SO₂R₁₁;

R_(b) is is selected from the group consisting of:

1) Hydrogen;

2) Halogen;

3) CF₃;

4) CN;

5) Substituted or unsubstituted —C₁-C₈ alkyl;

6) Substituted or unsubstituted —C₂-C₈ alkenyl;

7) Substituted or unsubstituted —C₂-C₈ alkynyl;

8) Substituted or unsubstituted —C₃-C₈ cycloalkyl;

9) Substituted or unsubstituted —C₃-C₈ heterocycloalkyl;

10) Substituted or unsubstituted aryl;

11) Substituted or unsubstituted heteroaryl;

12) Substituted or unsubstituted arylalkyl;

13) —C(O)R₁₁;

14) —C(O)OR₁₁;

15) —C(O)NR₈R₉; and

16) —C(O)N(R₈)SO₂R₁₁;

In certain embodiments, the invention provides a compound represented by Formula (I) as described above, or a pharmaceutically acceptable salt, hydrate, solvate, ester or prodrug thereof, wherein R₁, R₂, R₃, R₄, and R₅ are each hydrogen, R₆ is ethyl and ring A, R₇ and m are as previously defined.

A second embodiment of the invention is a compound represented by Formula II or a pharmaceutically acceptable salt, hydrate, solvate, ester or prodrug thereof,

wherein ring A, R₁, R₂, R₃, R₆, R₇ and m are as previously defined.

A third embodiment of the invention is a compound represented by Formula III or a pharmaceutically acceptable salt, hydrate, solvate, ester or prodrug thereof

wherein ring A, R₁, R₂, R₆, R₇ and m are as previously defined.

Illustrative structures of formula (III) can be represented by, but not limited to, formulas (III-1)˜(III-12), where R₆, R₇ and m are as previously defined:

A fourth embodiment of the invention is a compound represented by Formula IV or a pharmaceutically acceptable salt, solvate, hydrate, ester or prodrug thereof,

wherein R₇ and m are as previously defined.

Representative compounds of the invention include, but are not limited to, the following compounds (compound 1 to compound 87 in Table 1) according to Formula IV, wherein, R₇ and m are delineated for each compound in Table 1.

TABLE 1 compound m R₇ 1 0 H 2 0

3 0

4 0

5 0

6 0 —NH₂ 7 0 —NMe₂ 8 0

9 0

10 0

11 0

12 0

13 0

14 0

15 0

16 0

17 0

18 0

19 0

20 0

21 0

22 0

23 0

24 0

25 0

26 0

27 0

28 0

29 0

30 1 H 31 1

32 1

33 1

34 1

35 1 —NH₂ 36 1 —NMe₂ 37 1

38 1

39 1

40 1

41 1

42 1

43 1

44 1

45 1

46 1

47 1

48 1

49 1

50 1

51 1

52 1

53 1

54 1

55 1

56 1

57 1

58 1

59 2 H 60 2

61 2

62 2

63 2

64 2 —NH₂ 65 2 —NMe₂ 66 2

67 2

68 2

69 2

70 2

71 2

72 2

73 2

74 2

75 2

76 2

77 2

78 2

79 2

80 2

81 2

82 2

83 2

84 2

85 2

86 2

87 2

A fifth embodiment of the invention is a compound represented by Formula V or a pharmaceutically acceptable salt, solvate, hydrate, ester or prodrug thereof,

wherein R₇ and m are as previously defined.

Representative compounds of the invention include, but are not limited to, the following compounds (compound 88 to compound 174 in Table 2) according to Formula V, wherein, R₇ and m are delineated for each compound in Table 2.

TABLE 2 com- pound m R₇ 88 0 H 89 0

90 0

91 0

92 0

93 0 —NH₂ 94 0 —NMe₂ 95 0

96 0

97 0

98 0

99 0

100 0

101 0

102 0

103 0

104 0

105 0

106 0

107 0

108 0

109 0

110 0

111 0

112 0

113 0

114 0

115 0

116 0

117 1 H 118 1

119 1

120 1

121 1

122 1 —NH₂ 123 1 —NMe₂ 124 1

125 1

126 1

127 1

128 1

129 1

130 1

131 1

132 1

133 1

134 1

135 1

136 1

137 1

138 1

139 1

140 1

141 1

142 1

143 1

144 1

145 1

146 2 H 147 2

148 2

149 2

150 2

151 2 —NH₂ 152 2 —NMe₂ 153 2

154 2

155 2

156 2

157 2

158 2

159 2

160 2

161 2

162 2

163 2

164 2

165 2

166 2

167 2

168 2

169 2

170 2

171 2

172 2

173 2

174 2

A sixth embodiment of the invention is a compound represented by Formula VI or a pharmaceutically acceptable salt, solvate, hydrate, ester or prodrug thereof,

wherein R₇ and m are as previously defined.

Representative compounds of the invention include, but are not limited to, the following compounds (compound 175 to compound 261 in Table 3) according to Formula VI, wherein R₇ and m are delineated for each compound in Table 3.

TABLE 3 compound m R₇ 175 0 H 176 0

177 0

178 0

179 0

180 0 —NH₂ 181 0 —NMe₂ 182 0

183 0

184 0

185 0

186 0

187 0

188 0

189 0

190 0

191 0

192 0

193 0

194 0

195 0

196 0

197 0

198 0

199 0

200 0

201 0

202 0

203 0

204 1 H 205 1

206 1

207 1

208 1

209 1 —NH₂ 210 1 —NMe₂ 211 1

212 1

213 1

214 1

215 1

216 1

217 1

218 1

219 1

220 1

221 1

222 1

223 1

224 1

225 1

226 1

227 1

228 1

229 1

230 1

231 1

232 1

233 2 H 234 2

235 2

236 2

237 2

238 2 —NH₂ 239 2 —NMe₂ 240 2

241 2

242 2

243 2

244 2

245 2

246 2

247 2

248 2

249 2

250 2

251 2

252 2

253 2

254 2

255 2

256 2

257 2

258 2

259 2

260 2

261 2

A seventh embodiment of the invention is a compound represented by Formula VII or a pharmaceutically acceptable salt, solvate, hydrate, ester or prodrug thereof,

wherein R₇ and m are as previously defined.

Representative compounds of the invention include, but are not limited to, the following compounds (compound 262 to compound 348 in Table 4) according to Formula VII, wherein, R₇ and m are delineated for each compound in Table 4.

TABLE 4 compound m R₇ 262 0 H 263 0

264 0

265 0

266 0

267 0 —NH₂ 268 0 —NMe₂ 269 0

270 0

271 0

272 0

273 0

274 0

275 0

276 0

277 0

278 0

279 0

280 0

281 0

282 0

283 0

284 0

285 0

286 0

287 0

288 0

289 0

290 0

291 1 H 292 1

293 1

294 1

295 1

296 1 —NH₂ 297 1 —NMe₂ 298 1

299 1

300 1

301 1

302 1

303 1

304 1

305 1

306 1

307 1

308 1

309 1

310 1

311 1

312 1

313 1

314 1

315 1

316 1

317 1

318 1

319 1

320 2 H 321 2

322 2

323 2

324 2

325 2 —NH₂ 326 2 —NMe₂ 327 2

328 2

329 2

330 2

331 2

332 2

333 2

334 2

335 2

336 2

337 2

338 2

339 2

340 2

341 2

342 2

343 2

344 2

345 2

346 2

347 2

348 2

In another embodiment, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound or combination of compounds of the present invention, or a pharmaceutically acceptable salt form, stereoisomer, solvate, hydrate or combination thereof, in combination with a pharmaceutically acceptable carrier or excipient.

In yet another embodiment, the present invention provides a method for the prevention or treatment of a TGR5 mediated disease or condition. The method comprises administering a therapeutically effective amount of a compound of formula (I). The present invention also provides the use of a compound of formula (I) for the preparation of a medicament for the prevention or treatment of a TGR5 mediated disease or condition.

In certain embodiments, a disease that involves modulation of the TGR5 receptor is selected from metabolic disease, inflammatory disease, liver disease, autoimmune disease, cardiac disease, kidney disease, cancer, and gastrointestinal disease.

In certain embodiments, the present invention provides a method for the prevention or treatment of an FXR mediated disease or condition. The method comprises administering a therapeutically effective amount of a compound of formula (I). The present invention also provides the use of a compound of formula (I) for the preparation of a medicament for the prevention or treatment of an FXR mediated disease or condition.

In certain embodiments, the FXR-mediated disease or condition is cardiovascular disease, atherosclerosis, arteriosclerosis, hypercholesteremia, or hyperlipidemia chronic liver disease, gastrointestinal disease, renal disease, metabolic disease, cancer (i.e., colorectal cancer), or neurological indications such as stroke.

In certain embodiments, the chronic liver disease is primary biliary cirrhosis (PBC), cerebrotendinous xanthomatosis (CTX), primary sclerosing cholangitis (PSC), drug induced cholestasis, intrahepatic cholestasis of pregnancy, parenteral nutrition associated cholestasis (PNAC), bacterial overgrowth or sepsis associated cholestasis, autoimmune hepatitis, chronic viral hepatitis, alcoholic liver disease, nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), liver transplant associated graft versus host disease, living donor transplant liver regeneration, congenital hepatic fibrosis, choledocholithiasis, granulomatous liver disease, intra- or extrahepatic malignancy, Sjogren's syndrome, Sarcoidosis, Wilson's disease, Gaucher's disease, hemochromatosis, or alpha 1-antitrypsin deficiency. In certain embodiments, the gastrointestinal disease is inflammatory bowel disease (IBD) (including Crohn's disease and ulcerative colitis), irritable bowel syndrome (IBS), bacterial overgrowth, malabsorption, post-radiation colitis, or microscopic colitis.

In certain embodiments, the renal disease is diabetic nephropathy, focal segmental glomerulosclerosis (FSGS), hypertensive nephrosclerosis, chronic glomerulonephritis, chronic transplant glomerulopathy, chronic interstitial nephritis, or polycystic kidney disease.

In certain embodiments, the cardiovascular disease is atherosclerosis, arteriosclerosis, dyslipidemia, hypercholesterolemia, or hypertriglyceridemia.

In certain embodiments, the metabolic disease is insulin resistance, Type I or Type II diabetes, or obesity.

In yet another embodiment, the invention provides the use of the compound or pharmaceutical composition of the invention, in the manufacture of a medicament for a treating or preventing a disease in a subject that involves modulation of the TGR5 receptor. The invention includes a method of treating or preventing a disease that involves modulation of the TGR5 receptor in a subject by administering a compound or pharmaceutical composition of the invention.

In certain embodiments, a disease that involves modulation of the TGR5 receptor is selected from metabolic disease, inflammatory disease, liver disease, autoimmune disease, cardiac disease, kidney disease, cancer, and gastrointestinal disease.

In one aspect, the invention provides for the use, wherein the disease is an inflammatory disease selected from allergy, osteoarthritis, appendicitis, bronchial asthma, pancreatitis, allergic rash, and psoriasis. The invention includes a method of treating or preventing an inflammatory disease selected from allergy, osteoarthritis, appendicitis, bronchial asthma, pancreatitis, allergic rash, and psoriasis.

In one aspect, the invention provides for the use, wherein the disease is an autoimmune disease selected from rheumatoid arthritis, multiple sclerosis, and type I diabetes. The invention includes a method of treating or preventing an autoimmune disease selected from rheumatoid arthritis, multiple sclerosis, and type I diabetes.

In one aspect, the invention provides for the use, wherein the disease is a gastrointestinal disease selected from inflammatory bowel disease (Crohn's disease, ulcerative colitis), short bowel syndrome (post-radiation colitis), microscopic colitis, irritable bowel syndrome (malabsorption), and bacterial overgrowth. The invention includes a method of treating or preventing a gastrointestinal disease selected from inflammatory bowel disease (Crohn's disease, ulcerative colitis), short bowel syndrome (post-radiation colitis), microscopic colitis, irritable bowel syndrome (malabsorption), and bacterial overgrowth.

In one aspect, the invention provides for the use, wherein the disease is kidney disease selected from diabetic nephropathy, chronic renal failure, hypertensive nephrosclerosis, chronic glomerulonephritis, chronic transplant glomerulopathy, chronic interstitial nephritis, and polycystic kidney disease. The invention includes a method of treating or preventing kidney disease selected from diabetic nephropathy, chronic renal failure, hypertensive nephrosclerosis, chronic glomerulonephritis, chronic transplant glomerulopathy, chronic interstitial nephritis, and polycystic kidney disease.

In one aspect, the invention provides for the use, wherein the disease is cancer selected from colorectal cancer, liver cancer, hepatocellular carcinoma, cholangio carcinoma, renal cancer, gastric cancer, pancreatic cancer, prostate cancer, and insulanoma. The invention includes a method of treating or preventing cancer selected from colorectal cancer, liver cancer, hepatocellular carcinoma, cholangio carcinoma, renal cancer, gastric cancer, pancreatic cancer, prostate cancer, and insulanoma.

In one aspect, the compound is a selective FXR agonist over TGR5 activator.

In one aspect, the compound is a selective TGR5 agonist over FXR activator.

In one aspect, the compound is a dual agonist for both FXR and TGR5.

Yet a further aspect of the present invention is a process of making any of the compounds delineated herein employing any of the synthetic means delineated herein.

DEFINITIONS

Listed below are definitions of various terms used to describe this invention. These definitions apply to the terms as they are used throughout this specification and claims, unless otherwise limited in specific instances, either individually or as part of a larger group.

The term “alkyl”, as used herein, refers to a saturated, monovalent straight- or branched-chain hydrocarbon group. Preferred alkyl radicals include C₁-C₆ alkyl and C₁-C₈ alkyl radicals. Examples of C₁-C₆ alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, tent-butyl, neopentyl, n-hexyl groups; and examples of C₁-C₈ alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, neopentyl, n-hexyl, heptyl, and octyl groups.

The term “alkenyl”, as used herein, denote a monovalent group derived from a hydrocarbon moiety by the removal of a single hydrogen atom wherein the hydrocarbon moiety has at least one carbon-carbon double bond. Preferred alkenyl groups include C₂-C₆ alkenyl and C₂-C₈ alkenyl groups. Alkenyl groups include, but are not limited to, for example, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl, heptenyl, octenyl and the like.

The term “alkynyl”, as used herein, denotes a monovalent group derived from a hydrocarbon moiety by the removal of a single hydrogen atom wherein the hydrocarbon moiety has at least one carbon-carbon triple bond. Preferred alkynyl groups include C₂-C₆ alkynyl and C₂-C₈ alkynyl groups. Representative alkynyl groups include, but are not limited to, for example, ethynyl, 1-propynyl, 1-butynyl, heptynyl, octynyl and the like.

The term “carbocycle” refers to a saturated (e.g., “cycloalkyl”), partially saturated (e.g., “cycloalkenyl” or “cycloalkynyl”) or completely unsaturated (e.g., “aryl”) ring system containing zero heteroatom ring atoms. “Ring atoms” or “ring members” are the atoms bound together to form the ring or rings. Where a carbocycle group is a divalent moiety linking two other elements in a depicted chemical structure (such as Z in Formula I_(A)), the carbocycle group can be attached to the two other elements through any two substitutable ring atoms. A C₄-C₆ carbocycle has 4-6 ring atoms.

The term “cycloalkyl”, as used herein, denotes a monovalent group derived from a monocyclic or polycyclic saturated carbocyclic ring compound by the removal of a single hydrogen atom. Preferred cycloalkyl groups include C₃-C₈ cycloalkyl and C₃-C₁₂ cycloalkyl groups. Examples of C₃-C₈-cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopentyl and cyclooctyl; and examples of C₃-C₁₂-cycloalkyl include, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclo [2.2.1] heptyl, and bicyclo [2.2.2] octyl.

The term “cycloalkenyl” as used herein, denote a monovalent group derived from a monocyclic or polycyclic carbocyclic ring compound having at least one carbon-carbon double bond by the removal of a single hydrogen atom. Preferred cycloalkenyl groups include C₃-C₈ cycloalkenyl and C₃-C₁₂ cycloalkenyl groups. Examples of C₃-C₈-cycloalkenyl include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, and the like; and examples of C₃-C₁₂-cycloalkenyl include, but not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, and the like.

The term “aryl,” as used herein, refers to a mono- or bicyclic carbocyclic ring system having one or two aromatic rings including, but not limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, indenyl and the like.

The term “arylalkyl,” as used herein, refers to a C₁-C₃ alkyl or C₁-C₆ alkyl residue attached to an aryl ring. Examples include, but are not limited to, benzyl, phenethyl and the like.

The term “heteroaryl,” as used herein, refers to a mono-, bi-, or tri-cyclic aromatic radical or ring having from five to ten ring atoms of which at least one ring atom is selected from S, O and N; wherein any N or S contained within the ring may be optionally oxidized. Preferred heteroaryl groups are monocyclic or bicyclic. Heteroaryl groups include, but are not limited to, pyridinyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isooxazolyl, thiadiazolyl, oxadiazolyl, thiophenyl, furanyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzooxazolyl, quinoxalinyl, and the like.

The term “heteroarylalkyl,” as used herein, refers to a C₁-C₃ alkyl or C₁-C₆ alkyl residue residue attached to a heteroaryl ring. Examples include, but are not limited to, pyridinylmethyl, pyrimidinylethyl and the like.

The term “substituted” as used herein, refers to independent replacement of one, two, or three or more of the hydrogen atoms thereon with substituents including, but not limited to, deuterium, —F, —Cl, —Br, —I, —OH, protected hydroxy, —NO₂, —CN, —NH₂, N₃, protected amino, alkoxy, thioalkoxy, oxo, C₁-C₁₂-alkyl, C₂-C₁₂-alkenyl, C₂-C₁₂-alkynyl, C₃-C₁₂-cycloalkyl, -halo-C₁-C₁₂-alkyl, -halo-C₂-C₁₂-alkenyl, -halo-C₂-C₁₂-alkynyl, -halo-C₃-C₁₂-cycloalkyl, —NH—C₁-C₁₂-alkyl, —NH—C₂-C₁₂-alkenyl, —NH—C₂-C₁₂-alkynyl, —NH—C₃-C₁₂-cycloalkyl, —NH-aryl, —NH-heteroaryl, —NH-heterocycloalkyl, -dialkylamino, -diarylamino, -diheteroarylamino, —O—C₂-C₁₂-alkenyl, —O—C₂-C₁₂-alkynyl, —O—C₃-C₁₂-cycloalkyl, —O-aryl, —O-heteroaryl, —O-heterocycloalkyl, —C(O)—C₁-C₁₂-alkyl, —C(O)—C₂-C₁₂-alkenyl, —C(O)—C₂-C₁₂-alkynyl, —C(O)—C₃-C₁₂-cycloalkyl, —C(O)-aryl, —C(O)-heteroaryl, —C(O)-heterocycloalkyl, —CONH₂, —CONH—C₁-C₁₂-alkyl, —CONH—C₂-C₁₂-alkenyl, —CONH—C₂-C₁₂-alkynyl, —CONH—C₃-C₁₂-cycloalkyl, —CONH-aryl, —CONH-heteroaryl, -CONH-heterocycloalkyl, —OCO₂—C₁-alkyl, —OCO₂—C₂-C₁₂-alkenyl, —OCO₂—C₂-C₁₂-alkynyl, —OCO₂—C₃-C₁₂-cycloalkyl, —OCO₂-aryl, —OCO₂-heteroaryl, —OCO₂-heterocycloalkyl, —OCONH₂, —OCONH—C₁-C₁₂-alkyl, —OCONH—C₂-C₁₂-alkenyl, —OCONH—C₂-C₁₂-alkynyl, —OCONH—C₃-C₁₂-cycloalkyl, —OCONH-aryl, —OCONH-heteroaryl, —OCONH-heterocycloalkyl, —OCON(C₁-C₁₂-alkyl)₂, —NHC(O)—C₁-C₁₂-alkyl, —NHC(O)—C₂-C₁₂-alkenyl, —NHC(O)—C₂-C₁₂-alkynyl, —NHC(O)—C₃-C₁₂-cycloalkyl, —NHC(O)-aryl, —NHC(O)-heteroaryl, —NHC(O)-heterocycloalkyl, —NHCO₂—C₁-C₁₂-alkyl, —NHCO₂—C₂-C₁₂-alkenyl, —NHCO₂—C₂-C₁₂-alkynyl, —NHCO₂—C₃-C₁₂-cycloalkyl, —NHCO₂-aryl, —NHCO₂-heteroaryl, —NHCO₂-heterocycloalkyl, —NHC(O)NH₂, —NHC(O)NH—C₁-C₁₂-alkyl, —NHC(O)NH—C₂-C₁₂-alkenyl, —NHC(O)NH—C₂-C₁₂-alkynyl, —NHC(O)NH—C₃-C₁₂-cycloalkyl, —NHC(O)NH-aryl, —NHC(O)NH-heteroaryl, —NHC(O)NH-heterocycloalkyl, NHC(S)NH₂, —NHC(S)NH—C₁-C₁₂-alkyl, —NHC(S)NH—C₂-C₁₂-alkenyl, —NHC(S)NH—C₂-C₁₂-alkynyl, —NHC(S)NH—C₃-C₁₂-cycloalkyl, —NHC(S)NH-aryl, —NHC(S)NH-heteroaryl, —NHC(S)NH-heterocycloalkyl, —NHC(NH)NH₂, —NHC(NH)NH—C₁-C₁₂-alkyl, —NHC(NH)NH—C₂-C₁₂-alkenyl, —NHC(NH)NH—C₂-C₁₂-alkynyl, —NHC(NH)NH—C₃-C₁₂-cycloalkyl, —NHC(NH)NH-aryl, —NHC(NH)NH-heteroaryl, —NHC(NH)NH-heterocycloalkyl, —NHC(NH)—C₁-C₁₂-alkyl, —NHC(NH)—C₂-C₁₂-alkenyl, —NHC(NH)—C₂-C₁₂-alkynyl, —NHC(NH)—C₃-C₁₂-cycloalkyl, —NHC(NH)-aryl, —NHC(NH)-heteroaryl, —NHC(NH)-heterocycloalkyl, —C(NH)NH—C₁-C₁₂-alkyl, —C(NH)NH—C₂-C₁₂-alkenyl, —C(NH)NH—C₂-C₁₂-alkynyl, —C(NH)NH—C₃-C₁₂-cycloalkyl, —C(NH)NH-aryl, —C(NH)NH-heteroaryl, —C(NH)NH-heterocycloalkyl, —S(O)—C₁-C₁₂-alkyl, —S(O)—C₂-C₁₂-alkenyl, —S(O)—C₂-C₁₂-alkynyl, —S(O)—C₃-C₁₂-cycloalkyl, —S(O)-aryl, —S(O)-heteroaryl, —S(O)-heterocycloalkyl —SO₂NH₂, —SO₂NH—C₁-C₁₂-alkyl, —SO₂NH—C₂-C₁₂-alkenyl, —SO₂NH—C₂-C₁₂-alkynyl, —SO₂NH—C₃-C₁₂-cycloalkyl, —SO₂NH-aryl, —SO₂NH-heteroaryl, —SO₂NH-heterocycloalkyl, —NHSO₂—C₁-C₁₂-alkyl, —NHSO₂—C₂-C₁₂-alkenyl, —NHSO₂—C₂-C₁₂-alkynyl, —NHSO₂—C₃-C₁₂-cycloalkyl, —NHSO₂-aryl, —NHSO₂-heteroaryl, —NHSO₂-heterocycloalkyl, —CH₂NH₂, —CH₂SO₂CH₃, -aryl, -arylalkyl, -heteroaryl, -heteroarylalkyl, -heterocycloalkyl, —C₃-C₁₂-cycloalkyl, polyalkoxyalkyl, polyalkoxy, -methoxymethoxy, -methoxyethoxy, —SH, —S—C₁-C₁₂-alkyl, —S—C₂-C₁₂-alkenyl, —S—C₂-C₁₂-alkynyl, —S—C₃-C₁₂-cycloalkyl, —S-aryl, —S-heteroaryl, —S-heterocycloalkyl, methylthiomethyl, or -L′-R′, wherein L′ is C₁-C₆alkylene, C₂-C₆alkenylene or C₂-C₆alkynylene, and R′ is aryl, heteroaryl, heterocyclic, C₃-C₁₂cycloalkyl or C₃-C₂cycloalkenyl. It is understood that the aryls, heteroaryls, alkyls, and the like can be further substituted. In some cases, each substituent in a substituted moiety is additionally optionally substituted with one or more groups, each group being independently selected from —F, —Cl, —Br, —I, —OH, —NO₂, —CN, or —NH₂.

In accordance with the invention, any of the aryls, substituted aryls, heteroaryls and substituted heteroaryls described herein, can be any aromatic group. Aromatic groups can be substituted or unsubstituted.

It is understood that any alkyl, alkenyl, alkynyl, cycloalkyl and cycloalkenyl moiety described herein can also be an aliphatic group, an alicyclic group or a heterocyclic group. An “aliphatic group” is non-aromatic moiety that may contain any combination of carbon atoms, hydrogen atoms, halogen atoms, oxygen, nitrogen or other atoms, and optionally contain one or more units of unsaturation, e.g., double and/or triple bonds. An aliphatic group may be straight chained, branched or cyclic and preferably contains between about 1 and about 24 carbon atoms, more typically between about 1 and about 12 carbon atoms. In addition to aliphatic hydrocarbon groups, aliphatic groups include, for example, polyalkoxyalkyls, such as polyalkylene glycols, polyamines, and polyimines, for example. Such aliphatic groups may be further substituted. It is understood that aliphatic groups may be used in place of the alkyl, alkenyl, alkynyl, alkylene, alkenylene, and alkynylene groups described herein.

The term “alicyclic,” as used herein, denotes a monovalent group derived from a monocyclic or polycyclic saturated carbocyclic ring compound by the removal of a single hydrogen atom. Examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclo [2.2.1] heptyl, and bicyclo [2.2.2] octyl. Such alicyclic groups may be further substituted.

The terms “heterocyclic” or “heterocycloalkyl” can be used interchangeably and referred to a non-aromatic ring or a bi- or tri-cyclic group spiro, fused or bridged system, where (i) each ring system contains at least one heteroatom independently selected from oxygen, sulfur and nitrogen, (ii) each ring system can be saturated or unsaturated (iii) the nitrogen and sulfur heteroatoms may optionally be oxidized, (iv) the nitrogen heteroatom may optionally be quaternized, (v) any of the above rings may be fused to an aromatic ring, and (vi) the remaining ring atoms are carbon atoms which may be optionally oxo-substituted. Representative heterocycloalkyl groups include, but are not limited to, 1,3-dioxolane, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinonyl, 2-azabicyclo[2.2.1]heptyl, and tetrahydrofuryl. Such heterocyclic groups may be further substituted. Heteroaryl or heterocyclic groups can be C-attached or N-attached (where possible).

It will be apparent that in various embodiments of the invention, the substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, arylalkyl, heteroarylalkyl, and heterocycloalkyl are intended to be monovalent or divalent. Thus, alkylene, alkenylene, and alkynylene, cycloaklylene, cycloalkenylene, cycloalkynylene, arylalkylene, hetoerarylalkylene and heterocycloalkylene groups are to be included in the above definitions, and are applicable to provide the formulas herein with proper valency.

The term “hydroxy activating group”, as used herein, refers to a labile chemical moiety which is known in the art to activate a hydroxy group so that it will depart during synthetic procedures such as in a substitution or elimination reactions. Examples of hydroxy activating group include, but not limited to, mesylate, tosylate, triflate, p-nitrobenzoate, phosphonate and the like.

The term “activated hydroxy”, as used herein, refers to a hydroxy group activated with a hydroxy activating group, as defined above, including mesylate, tosylate, triflate, p-nitrobenzoate, phosphonate groups, for example.

The term “protected hydroxy,” as used herein, refers to a hydroxy group protected with a hydroxy protecting group, as defined above, including benzoyl, acetyl, trimethylsilyl, triethylsilyl, and methoxymethyl groups.

The term “hydroxy protecting group,” as used herein, refers to a labile chemical moiety which is known in the art to protect a hydroxy group against undesired reactions during synthetic procedures. After said synthetic procedure(s) the hydroxy protecting group as described herein may be selectively removed. Hydroxy protecting groups as known in the are described generally in T. H. Greene and P. G., S. M. Wuts, Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons, New York (1999). Examples of hydroxy protecting groups include benzyloxycarbonyl, 4-nitrobenzyloxycarbonyl, 4-bromobenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, methoxycarbonyl, tert-butoxycarbonyl, isopropoxycarbonyl, diphenylmethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, 2-(trimethylsilyl)ethoxycarbonyl, 2-furfuryloxycarbonyl, allyloxycarbonyl, acetyl, formyl, chloroacetyl, trifluoroacetyl, methoxyacetyl, phenoxyacetyl, benzoyl, methyl, t-butyl, 2,2,2-trichloroethyl, 2-trimethylsilyl ethyl, 1,1-dimethyl-2-propenyl, 3-methyl-3-butenyl, allyl, benzyl, para-methoxybenzyldiphenylmethyl, triphenylmethyl (trityl), tetrahydrofuryl, methoxymethyl, methylthiomethyl, benzyloxymethyl, 2,2,2-triehloroethoxymethyl, 2-(trimethylsilyl)ethoxymethyl, methanesulfonyl, para-toluenesulfonyl, trimethylsilyl, triethylsilyl, triisopropylsilyl, and the like. Preferred hydroxy protecting groups for the present invention are acetyl (Ac or —C(O)CH₃), benzoyl (Bz or —C(O)C₆H₅), and trimethylsilyl (TMS or —Si(CH₃)₃).

The terms “halo” and “halogen,” as used herein, refer to an atom selected from fluorine, chlorine, bromine and iodine.

In certain embodiments, the compounds of each formula herein are defined to include isotopically labeled compounds. An “isotopically labeled compound” is a compound in which at least one atomic position is enriched in a specific isotope of the designated element to a level which is significantly greater than the natural abundance of that isotope. For example, one or more hydrogen atom positions in a compound can be enriched with deuterium to a level which is significantly greater than the natural abundance of deuterium, for example, enrichment to a level of at least 1%, preferably at least 20% or at least 50%. Such a deuterated compound may, for example, be metabolized more slowly than its non-deuterated analog, and therefore exhibit a longer half-life when administered to a subject. Such compounds can synthesize using methods known in the art, for example by employing deuterated starting materials. Unless stated to the contrary, isotopically labeled compounds are pharmaceutically acceptable.

The compounds described herein contain one or more asymmetric centers and thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that may be defined, in terms of absolute stereochemistry, as (R)- or (S)-, or as (D)- or (L)- for amino acids. The present invention is meant to include all such possible isomers, as well as their racemic and optically pure forms. Optical isomers may be prepared from their respective optically active precursors by the procedures described above, or by resolving the racemic mixtures. The resolution can be carried out in the presence of a resolving agent, by chromatography or by repeated crystallization or by some combination of these techniques, which are known to those skilled in the art. Further details regarding resolutions can be found in Jacques, et al., Enantiomers, Racemates, and Resolutions (John Wiley & Sons, 1981). When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, and unless specified otherwise, it is intended that the compounds include both E and Z geometric isomers. Likewise, all tautomeric forms are also intended to be included. The configuration of any carbon-carbon double bond appearing herein is selected for convenience only and is not intended to designate a particular configuration unless the text so states; thus a carbon-carbon double bond depicted arbitrarily herein as trans may be cis, trans, or a mixture of the two in any proportion.

The term “subject” as used herein refers to a mammal. A subject therefore refers to, for example, dogs, cats, horses, cows, pigs, guinea pigs, and the like. Preferably the subject is a human. When the subject is a human, the subject may be referred to herein as a patient.

As used herein, the term “pharmaceutically acceptable salt” refers to those salts of the compounds formed by the process of the present invention which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art.

Berge, et al. describes pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66: 1-19 (1977). The salts can be prepared in situ during the final isolation and purification of the compounds of the invention, or separately by reacting the free base function with a suitable organic acid. Examples of pharmaceutically acceptable salts include, but are not limited to, nontoxic acid addition salts e.g., salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include, but are not limited to, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, alkyl having from 1 to 6 carbon atoms, sulfonate and aryl sulfonate.

Pharmaceutically acceptable salts can also be prepared by deprotonation of the parent compound with a suitable base, thereby forming the anionic conjugate base of the parent compound. In such salts the counter ion is a cation. Suitable cations include ammonium and metal cations, such as alkali metal cations, including Li⁺, Na⁺, K⁺ and Cs⁺, and alkaline earth metal cations, such as Mg²⁺ and Ca²⁺.

The term “amino protecting group,” as used herein, refers to a labile chemical moiety which is known in the art to protect an amino group against undesired reactions during synthetic procedures. After said synthetic procedure(s) the amino protecting group as described herein may be selectively removed. Amino protecting groups as known in the are described generally in T. H. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons, New York (1999). Examples of amino protecting groups include, but are not limited to, t-butoxycarbonyl, 9-fluorenylmethoxycarbonyl, benzyloxycarbonyl, and the like.

As used herein, the term “pharmaceutically acceptable ester” refers to esters of the compounds formed by the process of the present invention which hydrolyze in vivo and include those that break down readily in the human body to leave the parent compound or a salt thereof. Suitable ester groups include, for example, those derived from pharmaceutically acceptable aliphatic carboxylic acids, particularly alkanoic, alkenoic, cycloalkanoic and alkanedioic acids, in which each alkyl or alkenyl moiety advantageously has not more than 6 carbon atoms. Examples of particular esters include, but are not limited to, formates, acetates, propionates, butyrates, acrylates and ethyl succinates.

The term “pharmaceutically acceptable prodrugs” as used herein refers to those prodrugs of the compounds formed by the process of the present invention which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals with undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio, and effective for their intended use, as well as the zwitterionic forms, where possible, of the compounds of the present invention. “Prodrug”, as used herein means a compound, which is convertible in vivo by metabolic means (e.g. by hydrolysis) to afford any compound delineated by the formulae of the instant invention. Various forms of prodrugs are known in the art, for example, as discussed in Bundgaard, (ed.), Design of Prodrugs, Elsevier (1985); Widder, et al. (ed.), Methods in Enzymology, Vol. 4, Academic Press (1985); Krogsgaard-Larsen, et al., (ed). “Design and Application of Prodrugs, Textbook of Drug Design and Development, Chapter 5, 113-191 (1991); Bundgaard, et al., Journal of Drug Deliver Reviews, 8:1-38(1992); Bundgaard, J. of Pharmaceutical Sciences, 77:285 et seq. (1988); Higuchi and Stella (eds.) Prodrugs as Novel Drug Delivery Systems, American Chemical Society (1975); and Bernard Testa & Joachim Mayer, “Hydrolysis In Drug And Prodrug Metabolism: Chemistry, Biochemistry And Enzymology,” John Wiley and Sons, Ltd. (2002).

The term “treating”, as used herein, means relieving, lessening, reducing, eliminating, modulating, or ameliorating, i.e. causing regression of the disease state or condition. Treating can also include inhibiting, i.e. arresting the development, of a existing disease state or condition, and relieving or ameliorating, i.e. causing regression of an existing disease state or condition, for example when the disease state or condition may already be present.

The term “preventing”, as used herein means, to completely or almost completely stop a disease state or condition, from occurring in a patient or subject, especially when the patient or subject is predisposed to such or at risk of contracting a disease state or condition.

Additionally, the compounds of the present invention, for example, the salts of the compounds, can exist in either hydrated or unhydrated (the anhydrous) form or as solvates with other solvent molecules. Nonlimiting examples of hydrates include monohydrates, dihydrates, etc. Nonlimiting examples of solvates include ethanol solvates, acetone solvates, etc.

“Solvates” means solvent addition forms that contain either stoichiometric or non stoichiometric amounts of solvent. Some compounds have a tendency to trap a fixed molar ratio of solvent molecules in the crystalline solid state, thus forming a solvate. If the solvent is water the solvate formed is a hydrate, when the solvent is alcohol, the solvate formed is an alcoholate. Hydrates are formed by the combination of one or more molecules of water with one of the substances in which the water retains its molecular state as H₂O, such combination being able to form one or more hydrate.

As used herein, the term “analog” refers to a chemical compound that is structurally similar to another but differs slightly in composition (as in the replacement of one atom by an atom of a different element or in the presence of a particular functional group, or the replacement of one functional group by another functional group). Thus, an analog is a compound that is similar to or comparable in function and appearance to the reference compound.

The term “aprotic solvent,” as used herein, refers to a solvent that is relatively inert to proton activity, i.e., not acting as a proton-donor. Examples include, but are not limited to, hydrocarbons, such as hexane and toluene, for example, halogenated hydrocarbons, such as, for example, methylene chloride, ethylene chloride, chloroform, and the like, heterocyclic compounds, such as, for example, tetrahydrofuran and N-methylpyrrolidinone, and ethers such as diethyl ether, bis-methoxymethyl ether. Such solvents are well known to those skilled in the art, and individual solvents or mixtures thereof may be preferred for specific compounds and reaction conditions, depending upon such factors as the solubility of reagents, reactivity of reagents and preferred temperature ranges, for example. Further discussions of aprotic solvents may be found in organic chemistry textbooks or in specialized monographs, for example: Organic Solvents Physical Properties and Methods of Purification, 4th ed., edited by John A. Riddick et al., Vol. II, in the Techniques of Chemistry Series, John Wiley & Sons, NY, 1986.

The terms “protogenic organic solvent” or “protic solvent” as used herein, refer to a solvent that tends to provide protons, such as an alcohol, for example, methanol, ethanol, propanol, isopropanol, butanol, t-butanol, and the like. Such solvents are well known to those skilled in the art, and individual solvents or mixtures thereof may be preferred for specific compounds and reaction conditions, depending upon such factors as the solubility of reagents, reactivity of reagents and preferred temperature ranges, for example. Further discussions of protogenic solvents may be found in organic chemistry textbooks or in specialized monographs, for example: Organic Solvents Physical Properties and Methods of Purification, 4th ed., edited by John A. Riddick et al., Vol. II, in the Techniques of Chemistry Series, John Wiley & Sons, NY, 1986.

Combinations of substituents and variables envisioned by this invention are only those that result in the formation of stable compounds. The term “stable”, as used herein, refers to compounds which possess stability sufficient to allow manufacture and which maintains the integrity of the compound for a sufficient period of time to be useful for the purposes detailed herein (e.g., therapeutic or prophylactic administration to a subject).

The synthesized compounds can be separated from a reaction mixture and further purified by a method such as column chromatography, high pressure liquid chromatography, or recrystallization. Additionally, the various synthetic steps may be performed in an alternate sequence or order to give the desired compounds. In addition, the solvents, temperatures, reaction durations, etc. delineated herein are for purposes of illustration only and variation of the reaction conditions can produce the desired bridged macrocyclic products of the present invention. Synthetic chemistry transformations and protecting group methodologies (protection and deprotection) useful in synthesizing the compounds described herein include, for example, those described in R. Larock, Comprehensive Organic Transformations, VCH Publishers (1989); T. W. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 2d. Ed., John Wiley and Sons (1991); L. Fieser and M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995).

The compounds of this invention may be modified by appending various functionalities via synthetic means delineated herein to enhance selective biological properties. Such modifications include those which increase biological penetration into a given biological system (e.g., blood, lymphatic system, central nervous system), increase oral availability, increase solubility to allow administration by injection, alter metabolism and alter rate of excretion.

Pharmaceutical Compositions

The pharmaceutical compositions of the present invention comprise a therapeutically effective amount of a compound of the present invention formulated together with one or more pharmaceutically acceptable carriers. As used herein, the term “pharmaceutically acceptable carrier” means a non-toxic, inert solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type. Some examples of materials which can serve as pharmaceutically acceptable carriers are sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil; safflower oil; sesame oil; olive oil; corn oil and soybean oil; glycols; such a propylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol, and phosphate buffer solutions, as well as other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the composition, according to the judgment of the formulator. The pharmaceutical compositions of this invention can be administered to humans and other animals orally, rectally, parenterally, intracisternally, intravaginally, intraperitoneally, topically (as by powders, ointments, or drops), buccally, or as an oral or nasal spray.

The pharmaceutical compositions of this invention may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir, preferably by oral administration or administration by injection. The pharmaceutical compositions of this invention may contain any conventional non-toxic pharmaceutically-acceptable carriers, adjuvants or vehicles. In some cases, the pH of the formulation may be adjusted with pharmaceutically acceptable acids, bases or buffers to enhance the stability of the formulated compound or its delivery form. The term parenteral as used herein includes subcutaneous, intracutaneous, intravenous, intramuscular, intraarticular, intraarterial, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, U.S.P. and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

In order to prolong the effect of a drug, it is often desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution, which, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle. Injectable depot forms are made by forming microencapsule matrices of the drug in biodegradable polymers such as polylactide-polyglycolide. Depending upon the ratio of drug to polymer and the nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions which are compatible with body tissues.

Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the compounds of this invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or: a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants such as glycerol, d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.

The active compounds can also be in micro-encapsulated form with one or more excipients as noted above. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings and other coatings well known in the pharmaceutical formulating art. In such solid dosage forms the active compound may be admixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions which can be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound of this invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active component is admixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffers as may be required. Ophthalmic formulation, ear drops, eye ointments, powders and solutions are also contemplated as being within the scope of this invention.

The ointments, pastes, creams and gels may contain, in addition to an active compound of this invention, excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to the compounds of this invention, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays can additionally contain customary propellants such as chlorofluorohydrocarbons.

Transdermal patches have the added advantage of providing controlled delivery of a compound to the body. Such dosage forms can be made by dissolving or dispensing the compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by either providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel.

Unless otherwise defined, all technical and scientific terms used herein are accorded the meaning commonly known to one with ordinary skill in the art. All publications, patents, published patent applications, and other references mentioned herein are hereby incorporated by reference in their entirety.

Abbreviations

Abbreviations which have been used in the descriptions of the schemes and the examples that follow are:

ACN for acetonitrile;

BME for 2-mercaptoethanol;

BOP for benzotriazol-1-yloxy-tris(dimethylamino)phosphonium hexafluorophosphate;

Burgess reagent for methyl N-(triethylammoniumsulfonyl)carbamate

Bu₃SnN₃ for tributyltin azide;

BzCl for benzoyl chloride;

CDI for carbonyldiimidazole;

COD for cyclooctadiene;

DABCO for 1,4-diazabicyclo[2.2.2]octane;

DAST for diethylaminosulfur trifluoride;

DABCYL for 6-(N-4′-carboxy-4-(dimethylamino)azobenzene)-aminohexyl-1-O-(2-cyanoethyl)-(N,N-diisopropyl)-phosphoramidite;

DBU for 1,8-Diazabicycloundec-7-ene;

DCC for N,N′-dicyclohexylcarbodiimide;

DCM for dichloromethane;

DIAD for diisopropyl azodicarboxylate;

DIBAL-H for diisobutylaluminum hydride;

DIPEA for diisopropyl ethylamine;

DMAP for N,N-dimethylaminopyridine;

DME for ethylene glycol dimethyl ether;

DMEM for Dulbecco's Modified Eagles Media;

DMF for N,N-dimethyl formamide;

DMSO for dimethylsulfoxide;

DSC for N,N′-disuccinimidyl carbonate;

DPPA for diphenylphosphoryl azide;

DUPHOS for

EDANS for 5-(2-Amino-ethylamino)-naphthalene-1-sulfonic acid;

EDCI or EDC for 1-(3-diethylaminopropyl)-3-ethylcarbodiimide hydrochloride;

Et₂O for diethyl ether;

EtOAc for ethyl acetate;

EtOH for ethyl alcohol;

HATU for O (7-Azabenzotriazole-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate;

HCl for hydrochloric acid;

Hoveyda's Cat. for Dichloro(o-isopropoxyphenylmethylene) (tricyclohexylphosphine)ruthenium(II);

In for indium;

KHMDS for potassium bis(trimethylsilyl) amide;

LAH for lithium aluminium hydride;

Lawesson's reagent for 2,4-Bis(4-methoxyphenyl)-1,3,2,4-dithiadiphosphetane-2,4-disulfide;

Ms for mesyl;

NaCN for sodium cyanide;

NaN₃ for sodium aizde;

NMM for N-4-methylmorpholine;

NMI for N-methylimidazole;

NMO for N-4-methylmorpholine-N-Oxide;

PyBrOP for Bromo-tri-pyrolidino-phosphonium hexafluorophosphate;

Ph for phenyl;

RCM for ring-closing metathesis;

RT for reverse transcription;

RT-PCR for reverse transcription-polymerase chain reaction;

T3P for propylphosphonic anhydride;

TBME for tert-butyl methyl ether;

TEA for triethyl amine;

Tf₂O for trifluoromethanesulfonic anhydride

TFA for trifluoroacetic acid;

THF for tetrahydrofuran;

TLC for thin layer chromatography;

(TMS)₂NH for hexamethyldisilazane

TMSOTf for trimethylsilyl trifluoromethanesulfonate;

TMSN₃ for trimethylsilyl azide;

TBS for t-Butyldimethylsilyl;

TMS for trimethylsilyl;

TPAP tetrapropylammonium perruthenate;

TPP or PPh₃ for triphenylphosphine;

TrCl for trityl chloride;

DMTrCl for 4,4′-dimethoxytrityl chloride;

tBOC or Boc for tert-butyloxy carbonyl.

Synthetic Methods

The compounds and processes of the present invention will be better understood in connection with the following synthetic schemes that illustrate the methods by which the compounds of the invention may be prepared, which are intended as an illustration only and not to limit the scope of the invention. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art and such changes and modifications including, without limitation, those relating to the chemical structures, substituents, derivatives, and/or methods of the invention may be made without departing from the spirit of the invention and the scope of the appended claims.

As shown in Scheme 1, novel bile acid analogs of the compound of formula (1-5) are prepared from the compound of formula (1-1), wherein m and R₆ are defined as previously, P is hydroxyl protecting groups. Thus, the indicated hydroxyl group of the compound of formula (1-1) are protected with P group to afford the compound of formula (1-2). P can be any hydroxyl protecting group such as, but not limited to Ac, Bz, chloroacetyl, TES, TBS, MOM and Bn. A more detailed discussion of the procedures, reagents and conditions for protection of hydroxyl group is described in literature, for example, by T. W. Greene and P. G. M. Wuts in “Protective Groups in Organic Synthesis” 3^(rd) ed., John Wiley & Son, Inc., 1999. Then, the compound of formula (1-2) is converted to the mixed anhydride compound of formula (1-3) using suitable reagent such as ClC(O)OR′, wherein R′ is C₁-C₆-alkyl, for example, isopropyl chloroformate. Further condensation of the compound of formula (1-3) with hydrazinecarboximidamide at elevated temperature affords the amino-triazole compound of formula (1-4). Then deprotection of P groups affords the compound of formula (1-5). A more detailed discussion of the procedures, reagents and conditions for deprotection of hydroxyl protecting groups is described in the literature, for example, by T. W. Greene and P. G. M. Wuts in “Protective Groups in Organic Synthesis” 3^(rd) ed., John Wiley & Son, Inc., 1999. A more detailed discussion of the procedures, reagents and conditions for such triazole synthesis is described in literature, for example, by N. V. Palysaeva and co-workers in “A Direct Approach to a 6-Hetarylamino[1,2,4]triazolo[4,3-b][1,2,4,5]tetrazine Library” from Organic Letters, 16(2), 406-409; 2014.

Scheme 2 illustrates the preparation of the amino-oxadiazo compound of formula (2-3) from the compound of formula (1-2), wherein m and R₆ are defined as previously, P is a hydroxyl protecting group. Thus, condensation of compound of formula (2-1) with cyanic bromide at elevated temperature affords the amino-oxadiazo compound of formula (2-2). Then deprotection of P groups affords the compound of formula (2-3). A more detailed discussion of the procedures, reagents and conditions for deprotection of hydroxyl protecting groups is described in the literature, for example, by T. W. Greene and P. G. M. Wuts in “Protective Groups in Organic Synthesis” 3^(rd) ed., John Wiley & Son, Inc., 1999. A more detailed discussion of the procedures, reagents and conditions for such amino-oxadiazo compound synthesis is described in literature, for example, by L. Chu and co-workers in “Evaluation of endo- and exo-aryl-substitutions and central scaffold modifications on diphenyl substituted alkanes as 5-lipoxygenase activating protein inhibitors” from Bioorganic & Medicinal Chemistry Letters (2012), 22, (12), 4133-4138.

Scheme 3 illustrates the preparation of the sulfonamide compound of formula (3-2) from the compound of formula (1-4), wherein m and R₆ are defined as previously, P is hydroxyl protecting groups. The compound of formula (1-4) is converted to the compound of formula (3-1) using sulfonyl chloride ClS(O)₂R₁₁, where R₁₁ is as previously defined. Then deprotection of P groups affords the compound of formula (3-2). A more detailed discussion of the procedures, reagents and conditions for deprotection of hydroxyl protecting groups is described in literature, for example, by T. W. Greene and P. G. M. Wuts in “Protective Groups in Organic Synthesis” 3^(rd) ed., John Wiley & Son, Inc., 1999.

Scheme 4 illustrates the preparation of the amide compound of formula (4-2) from the compound of formula (1-4), wherein m and R₆ are defined as previously, P is hydroxyl protecting groups. The compound of formula (1-4) is coupled with carboxylic acid R₁₁C(O)OH, where R₁₁ is as previously defined, using suitable coupling condition to give the compound of formula (4-1). The coupling reagent can be selected from, but not limited to, DCC, EDC, CDI, di-isopropyl carbodiimide, BOP-Cl, PyBOP, PyAOP, TFFH and HATU. Suitable bases include, but are not limited to, triethylamine, diisopropylethylamine, DBU, N-methylmorpholine and DMAP. The coupling reaction is carried out in an aprotic solvent such as, but not limited to, CH₂Cl₂, DMF or THF. The reaction temperature can vary from 0° C. to about 50° C. Then deprotection of P groups affords the compound of formula (4-2). A more detailed discussion of the procedures, reagents and conditions for deprotection of hydroxyl protecting groups is described in literature, for example, by T. W. Greene and P. G. M. Wuts in “Protective Groups in Organic Synthesis” 3^(rd) ed., John Wiley & Son, Inc., 1999.

Scheme 5 illustrates the preparation of the carbamate compound of formula (5-2) from the compound of formula (1-4), wherein m and R₆ are defined as previously, P is hydroxyl protecting groups. The compound of formula (1-4) is converted to the compound of formula (5-1) using suitable reagent such as, but not limited to, CDI and alcohol R₁₁OH, where R₁₁ is as previously defined. Then deprotection of P groups affords the compound of formula (4-2). A more detailed discussion of the procedures, reagents and conditions for deprotection of hydroxyl protecting groups is described in literature, for example, by T. W. Greene and P. G. M. Wuts in “Protective Groups in Organic Synthesis” 3^(rd) ed., John Wiley & Son, Inc., 1999.

Scheme 6 illustrates the preparation of the oxadiazo compound of formula (6-4) from the compound of formula (1-1), wherein m and R₆ are defined as previously, P₁ and P₂ are hydroxyl protecting groups. Thus, the two hydroxyl groups of the compound of formula (1-1) are protected with P₁ and P₂ groups to afford the compound of formula (6-1). P₁ and P₂ can be same or different. P₁ and P₂ can be any hydroxyl protecting group such as, but not limited to Ac, Bz, chloroacetyl, TES, TBS, MOM and Bn. The compound of formula (6-1) is converted to the compound of formula (6-2) by coupled with acylhydrazide. Further condensation of compound of formula (6-2) in the presence of Burgess reagent at elevated temperature affords oxadiazo compound of formula (6-3). Then deprotection of P₁and P₂ groups affords the compound of formula (6-4). A more detailed discussion of the procedures, reagents and conditions for deprotection of hydroxyl protecting groups is described in literature, for example, by T. W. Greene and P. G. M. Wuts in “Protective Groups in Organic Synthesis” 3^(rd) ed., John Wiley & Son, Inc., 1999. A more detailed discussion of the procedures, reagents and conditions for such 1,3,4-oxadiazo compound synthesis is described in literature, for example, by C. Li and co-workers in “A mild, one-pot preparation of 1,3,4-oxadiazoles” from Tetrahedron Letters, 50(47), 6435-6439; 2009.

Scheme 7 illustrates the preparation of the thiadiazo compound of formula (7-2) from the compound of formula (6-2), wherein m and R₆ are defined as previously, P₁and P₂ are hydroxyl protecting groups. Further condensation of compound of formula (6-2) in the presence of Lawesson's reagent at elevated temperature affords thiadiazo compound of formula (7-1). Then deprotection of Pand P₂ groups affords the compound of formula (7-2). A more detailed discussion of the procedures, reagents and conditions for deprotection of hydroxyl protecting groups is described in literature, for example, by T. W. Greene and P. G. M. Wuts in “Protective Groups in Organic Synthesis” 3^(rd) ed., John Wiley & Son, Inc., 1999. A more detailed discussion of the procedures, reagents and conditions for such 1,3,4-thiadiazo compound synthesis is described in literature, for example, by J. K. Augustine and co-workers in “Propylphosphonic anhydride (T3P): an efficient reagent for the one-pot synthesis of 1,2,4-oxadiazoles, 1,3,4-oxadiazoles, and 1, 3,4-thiadiazoles” from Tetrahedron (2009), 65(48), 9989-9996.

Scheme 8 illustrates the preparation of the compound of formula (8-2) from the compound of formula (1-2), wherein m and R₆ are defined as previously, P is hydroxyl protecting groups. Further condensation of compound of formula (1-2) with hydroxyacetimidamide at elevated temperature affords compound of formula (8-1). Then deprotection of P group affords the compound of formula (8-2). A more detailed discussion of the procedures, reagents and conditions for deprotection of hydroxyl protecting groups is described in literature, for example, by T. W. Greene and P. G. M. Wuts in “Protective Groups in Organic Synthesis” 3^(rd) ed., John Wiley & Son, Inc., 1999. A more detailed discussion of the procedures, reagents and conditions for such 1,2,4-oxadiazo compound synthesis is described in literature, for example, by J. K. Augustine and co-workers in “Propylphosphonic anhydride (T3P): an efficient reagent for the one-pot synthesis of 1,2,4-oxadiazoles, 1,3,4-oxadiazoles, and 1,3,4-thiadiazoles” from Tetrahedron (2009), 65(48), 9989-9996.

Scheme 9 illustrates the preparation of the triazole compound of formula (9-2) from the compound of formula (2-1), wherein m and R₆ are defined as previously, P is hydroxyl protecting groups. Further condensation of compound of formula (1-2) with nitrile compound at elevated temperature affords compound of formula (9-1). Then deprotection of P group affords the triazole compound of formula (9-2). A more detailed discussion of the procedures, reagents and conditions for deprotection of hydroxyl protecting groups is described in literature, for example, by T. W. Greene and P. G. M. Wuts in “Protective Groups in Organic Synthesis” 3^(rd) ed., John Wiley & Son, Inc., 1999.

EXAMPLES

The compounds and processes of the present invention will be better understood in connection with the following examples, which are intended as an illustration only and not limiting of the scope of the invention. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art and such changes and modifications including, without limitation, those relating to the chemical structures, substituents, derivatives, formulations and/or methods of the invention may be made without departing from the spirit of the invention and the scope of the appended claims.

Example 1

Step 1-1:

To a 1-dram vial were added 6(a)-ethyl-chenodeoxycholic acid (6-ECDCA) (100 mg, 0.24 mmol), anhydrous THF (3 mL), TBSC1 (107.5 mg, 0.71 mmol), and imidazole (57.2 mg, 0.84 mmol) respectively and the reaction mixture was stirred at rt for 22 h. Transferred to a separation funnel, diluted with EtOAC (50 mL) and washed with brine-water (10 mL, 1:1 v/v). Dried, filtered and concentrated and the resulting white solid was dissolved in MeOH (10 mL) and was added K₂CO₃ (49.7 mg, 0.36 mmol). The mixture was stirred at room temperature for 30 min. and cooled to 0° C. Then the reaction mixture was acidified by addition of 0.1 NHCl to PH<7 and diluted with EtOAc (30 mL). After washed with brine (10 mL), dried, and concentrated, the resulting crude material purified by CombiFlash (12 g SiO₂, acetone/hexanes=0−>40%) to give the compound (1) as a white solid, 88 mg, 65.6% yield over 2 steps.

Step 1-2:

To a solution of carboxylic acid compound (1) (536.1 mg, 1.0 mmol) in CH₂Cl₂ (20 mL) was added HOBT (142.2 mg, 1.05 mmol) and DCC (217.1 mg, 1.05 mmol). The reaction mixture was allowed to stir at room temperature for 30 min before addition of benzyl carbazate (174.9 mg, 1.05 mmol). Then the reaction was allowed to stir for overnight. The crude was filtered and further washed with EtOAc. The filtrate was then concentrated. The resulting crude was purified by CombiFlash (20 g SiO₂, EtOAc/hexanes=0->50%) to give the compound (2) as a white solid, 602.6 mg, 88% yield.

Step 1-3:

To a solution of compound (2) (602.6 mg, 0.882 mmol) in EtOH (9 mL) was added Pd/C (60 mg). The resulting suspension was purged with H2 three times. The reaction was then allowed to stir at room temperature under H₂ atmosphere for overnight. The crude was filtered through a short SiO₂ plug and washed with EtOAc. The filtrate was concentrated to afford a while solid as compound (3), which was directly used in the following transformation without any further purification, 482 mg, quant. yield.

Step 1-4:

To a solution of compound (3) (125.4 mg, 0.228 mmol) in n-BuOH (1 mL) was added cyclopropanecarbonitrile (0.254 mL, 3.43 mmol) and K₂CO₃ (47.3 mg, 0.342 mmol). The resulting suspension was allowed to heat in microwave reactor at 165° C. for 30 min. TLC indicated not full completion. Then the reaction was further heated in oil batch at 120° C. for overnight. The crude was filtered through a cotton plug. The filtrate was concentrated and purified by CombiFlash (12 g SiO₂, EtOAc/hexanes=0->50%) to give the compound (4) as a white solid, 52 mg, 38% yield.

Step 1-5:

To a solution of compound (4) (52 mg, 0.087 mmol) in MeOH (1 mL) was added 1 drop of aq. 37% HCl. The reaction was allowed to stir at room temperature for 10 min before concentration. The crude was purified by CombiFlash (12 g SiO₂, MeOH/DCM=0->10%) to give the triazole compound of example 1 as a white solid, 27 mg, 64% yield.

Example 2

Step 2-1:

To a solution of compound (1) (3.5 g, 6.543 mmol) in DCM (100 mL) at 0° C. was added N-methyl imidazole (2.08 mL, 26.17 mmol), bis(trimethylsilyl)acetamide (8 mL, 32.72 mmol) and TMSCl (2.5 mL, 19.63 mmol) slowly. The reaction was allowed to stir at room temperature for 2 hours before concentration. The residue was then redissolved in EtOAc and washed with 10% aq. citric acid solution. The organic layer was dried by Na₂SO₄, filtered and concentrated. The crude was then purified by CombiFlash (80 g SiO₂, EtOAc/hexanes=0->50%) to give the carboxylic compound (5) as a white solid, 3.45 g, 87% yield.

Step 2-2:

To a solution of compound (5) (91.8 mg, 0.151 mmol) in EtOAc (2 mL) was added pivalohydrazide (18.4 mg, 0.159 mmol), triethyl amine (53 μL, 0.378 mmol), Lawesson's reagent (92 mg, 0.227 mmol) and propylphosphonic anhydride (0.14 mL 50wt % EtOAc solution, 0.227 mmol) at room temperature. The reaction was allowed to stir at 80° C. for 8 hours before quenched by aq. NaHCO₃ at room temperature. The mixture was further extracted by EtOAc (25 mL×3). The combined organic layers were dried over Na₂SO₄, filtered and concentrated. The crude was then purified by CombiFlash (12 g SiO₂, EtOAc/hexanes=0->30%) to give the thiadiazo compound (6) as a white solid, 53.4 mg, 50% yield.

Step 2-3:

To a solution of compound (6) (53.4 mg, 0.076 mmol) in MeOH/THF (1 mL/0.5 mL) was added 1 drop of aq. 37% HCl. The reaction was allowed to stir at room temperature for 10 min before concentration. The crude was purified by CombiFlash (4 g SiO₂, EtOAc/hexanes=0->100%) to give the thiadiazo compound of example 2 as a white solid, 31.3 mg, 80% yield.

Example 3

Step 3-1:

To a solution of compound (5) (259 mg, 0.427 mmol) in 2 mL CH₂Cl₂ (4 mL) was charged cyclopropanecarbohydrazide (64.1 mg, 0.64 mmol), EDCI (163.7 mg, 0.854 mmol) and DMAP (104.3 mg, 0.854 mmol). The resulting solution was allowed to stir at room temperature until completion indicated by TLC. The solvent was removed under reduced pressure. The crude was redissolved in EtOAc and washed with 10% aq. citric acid. The aq. layer was then extracted with ethyl acetate (30 mL×2). The combined organic layers were dried over Na₂SO₄, filtered and concentrated under reduced pressure. The residue was purified by CombiFlash (20 g SiO₂, EtOAc/hexanes=0->50%) to give the compound (7) as a white solid, 217.8 mg, 74% yield.

Step 3-2:

To a solution of compound (7) (42.7 mg, 0.062 mmol) in THF (1 mL) was added Burgess reagent (29.5 mg, 0.124 mmol). The reaction was allowed to heat in microwave reactor at 140° C. for 15 min. The crude was concentrated and purified by CombiFlash (4 g SiO₂, EtOAc/hexanes=0->50%) to give the oxadiazo compound (8) as a white solid, 16.7 mg, 40% yield.

Step 3-3:

To a solution of compound (8) (34.9 mg, 0.052 mmol) in MeOH (1 mL) was added 1 drop of aq. 37% HCl. The reaction was allowed to stir at room temperature for 10 min before concentration. The crude was purified by CombiFlash (4 g SiO₂, MeOH/DCM=0->10%) to give the oxadiazo compound of example 3 as a white solid, 11.1 mg, 44% yield.

Example 4

Step 4-1:

To a solution of carboxylic acid compound (5) (106.6 mg, 0.176 mmol) in THF (2 mL) was added HOBT (24.9 mg, 0.184 mmol) and DCC (38 mg, 0.184 mmol). The reaction mixture was allowed to stir at room temperature for 30 min before addition of N-hydroxycyclopropanecarboximidamide (18.5 mg, 0.184 mmol). Then the reaction was allowed to stir for overnight. The crude was filtered and further washed with EtOAc. Then the resulting solution was concentrated and redissolved in dioxane (1 mL) and charged with triethyl amine (0.122 mL, 0.878 mmol). The reaction mixture was then allowed to heat in microwave reactor at 160° C. for 30 min. The crude was concentrated and purified by CombiFlash (12 g SiO₂, EtOAc/hexanes=0->30%) to give the compound (9) as a colorless oil, 41.2 mg, 35% yield over 2 steps.

Step 4-2:

To a solution of compound (9) (41.2 mg, 0.061 mmol) in MeOH (1 mL) was added 1 drop of aq. 37% HCl. The reaction was allowed to stir at room temperature for 10 min before concentration. The crude was purified by CombiFlash (4 g SiO₂, MeOH/DCM=0->10%) to give the oxadiazo compound of example 4 as a white solid, 28.2 mg, 95% yield.

Example 5

Step 5-1:

To a solution of compound (3) (394.7 mg, 0.719 mmol) in dioxane/water (7 mL/2.1 mL) was added NaHCO₃ (60.4 mg, 0.719 mmol) and cyanic bromide (76.2 mg, 0.719 mmol). The reaction was allowed to stir at room temperature for overnight. The reaction mixture was then concentrated and purified by CombiFlash (24 g SiO₂, MeOH/DCM=0->10%) to give the compound (10) as a white solid, 97.3 mg, 24% yield.

Step 5-2:

To a solution of compound (10) (38 mg, 0.0662 mmol) in MeOH (1 mL) was added 1 drop of aq. 37% HCl. The reaction was allowed to stir at room temperature for 10 min before concentration. The crude was purified by CombiFlash (4 g SiO₂, MeOH/DCM=0->10%) to give the amino-oxadiazo compound of example 5 as a white solid, 10.9 mg, 36% yield.

Example 6

Step 6-1:

To a solution of carboxylic acid (1) (1.493 g, 2.79 mmol) in THF (14 mL) was triethyl amine (1.56 mL, 11.2 mmol) and isopropyl chloroformate (6.14 mL 1.0 M PhMe solution, 6.14 mmol) slowly at 0° C. The reaction was allowed to warm to room temperature and stir for 1 hour. The solvent was removed under reduced pressure. The crude was redissolved in EtOAc and washed with water. The aq. layer was then extracted with ethyl acetate (30 mL×2). The combined organic layers were dried over Na₂SO₄, filtered and concentrated under reduced pressure. The crude oil of compound (11) was directly used in the following transformation without any further purification.

Step 6-2:

To a solution of of compound (11) obtained from previous step in toluene (6 mL) was added hydrazinecarboximidamide bicarbonate (296.4 mg, 2.18 mmol). The reaction was allowed to stir at 110° C. for 20 hours. The crude was filtered and purified by CombiFlash (20 g SiO₂, MeOH/DCM=0->10%) to give the amino-triazole compound (12) as a white solid, 166 mg, 27% yield over steps.

Step 6-3:

To a solution of compound (12) (19.1 mg, 0.033 mmol) in MeOH (1 mL) was added 1 drop of aq. 37% HCl. The reaction was allowed to stir at room temperature for 10 min before concentration. The crude was purified by CombiFlash (4 g SiO₂, MeOH/DCM=0->20%) to give the amino-triazole compound of example 6 as a white solid, 7.5 mg, 49% yield.

Example 7

Step 7-1:

To a solution of compound (12) (49 mg, 0.0856 mmol) in DCM (1 mL) was added methanesulfonyl chloride (8 μL, 0.1026 mmol) and triethyl amine (36 μL, 0.2568 mmol) at 0° C. The reaction was allowed to stir at room temperature for overnight. The solvent was removed under reduced pressure. The crude was redissolved in MeOH (1 mL) and charged with 1 drop of aq. 37% HCl. The reaction was allowed to stir at room temperature for 10 min before concentration. The crude was purified by CombiFlash (4 g SiO₂, MeOH/DCM=0->20%) to give the sulfonamide compound of example 7 as a white solid, 24.3 mg, 53% yield over 2 steps.

TABLE 1 Example structure HRMS: ¹H NMR 1

699.35 (M + HCO₂ ⁻) 2

605.41 (M + 1) 3

459.37 (M + 1) 4

537.35 (M + 1) 5

563.35 (M + 1) 1H NMR (500 MHz, Chloroform-d) d 5.89 (s, 2H), 3.70 (s, 1H), 3.41 (s, 1H, br), 2.82 (s, 1H), 2.63 (d, J = 12.1 Hz, 1H), 2.46 (s, 1H), 1.96-1.72 (m, 10H), 1.71-1.55 (m, 2H), 1.54-1.37 (m, 7H), 1.34-1.08 (m, 8H), 0.98-90 (m, 5H), 0.89 (s, 6H), 0.65 (s, 3H). 6

566.37 (M + 1) 7

635.38 (M + HCO₂ ⁻) 8

596.35 (M + HCO₂ ⁻) 9

460.35 (M + 1) 10

485.38 (M + 1) 11

467.37 (M + 1 − H₂O) 12

501.35 (M + 1) 13

484.40 (M + 1) 14

488.39 (M + 1) 15

537.35 (M + 1) 16

551.37 (M + 1) 17

517.39 (M + 1) 18

587.39 (M + HCO₂ ⁻) 19

637.40 (M + HCO₂ ⁻) 20

563.42 (M + 1)

Assays Human FXR (NR1H4) Assay

Determination of a ligand mediated Gal4 promoter driven transactivation to quantify ligand binding mediated activation of FXR. FXR Reporter Assay kit purchased from Indigo Bioscience (Catalogue number: IB00601) to determine the potency and efficacy of compound developed by Enanta that can induce FXR activation. The principal application of this reporter assay system is to quantify functional activity of human FXR. The assay utilizes non-human mammalian cells, CHO (Chinese hamster ovary) cells engineered to express human NR1H4 protein (referred to as FXR). Reporter cells also incorporate the cDNA encoding beetle luciferase which catalyzes the substrates and yields photon emission. Luminescence intensity of the reaction is quantified using a plate-reading luminometer, Envision. Reporter Cells include the luciferase reporter gene functionally linked to an FXR responsive promoter. Thus, quantifying changes in luciferase expression in the treated reporter cells provides a sensitive surrogate measure of the changes in FXR activity. EC₅₀ and efficacy (normalize to CDCA set as 100%) is determined by XLFit. The assay is according to the manufacturer's instructions. In brief, the assay was performed in white, 96 well plates using final volume of 100 ul containing cells with different doses of compounds. Retrieve Reporter Cells from −80° C. storage. Perform a rapid thaw of the frozen cells by transferring a 10 ml volume of 37° C. cell recovery medium into the tube of frozen cells. Recap the tube of Reporter Cells and immediately place it in a 37° C. water bath for 5-10 minutes. Retrieve the tube of Reporter Cell Suspension from the water bath. Sanitize the outside surface of the tube with a 70% alcohol swab, and then transfer it into the cell culture hood. Dispense 90 μl of cell suspension into each well of the 96-well Assay Plate. Transfer the plate into 37° C. incubator, allowing the cells adherent to the bottom of the well. Dilute compounds in Dilution Plate (DP), and administrate to cells at Assay Plate (AP). DMSO content of the samples was kept at 0.2%. Cells were incubated for additional 22 hours before luciferase activities were measured. Thirty minutes before intending to quantify FXR activity, remove Detection Substrate and Detection Buffer from the refrigerator and place them in a low-light area so that they may equilibrate to room temperature. Remove the plate's lid and discard all media contents by ejecting it into an appropriate waste container. Gently tap the inverted plate onto a clean absorbent paper towel to remove residual droplets. Cells will remain tightly adhered to well bottoms. Add 100 μl of luciferase detection reagent to each well of the assay plate. Allow the assay plate to rest at room temperature for at least 5 minutes following the addition of LDR. Set the instrument (Envision) to perform a single 5 second “plate shake” prior to reading the first assay well. Read time may be 0.5 second (500 mSec) per well. EC₅₀ and Efficacy (normalize to CDCA set as 100%) is determined by XLFit.

In Vitro Human TGR5 (GPBAR1) Activity Assay

The potency and efficacy of the compounds of the invention on TGR5 receptor can be evaluated using in vitro assays which carried out using the express kit from DiscoverX (cAMP Hunter™ eXpress GPBAR1 CHO-K1 GPCR Assay; Catalogue number: 95-0049E2CP2S)GPBAR1 (G protein-coupled bile acid receptor 1) encodes a member of the G protein-coupled receptor (GPCR) superfamily. GPBAR1 activation following ligand binding initiates a series of second messenger cascades that result in a cellular response. Treatment of CHO cells expressing GPBAR1 with bile acids induces the production of intracellular cAMP and internalization of the receptor. The potency and efficacy of compound for GPBAR1 activation by measuring cyclic adenosine monophosphate (cyclic AMP or cAMP) levels in live cells using a competitive immunoassay based on Enzyme Fragment Complementation (EFC).

In brief, following seeding the cells into the white, 96 well microplate, place it in a 37° C., 5% CO2 in a humidified incubator for 18-24 hours prior to testing. On second day, proceed to the appropriate cAMP Hunter eXpress Protocol according to the manufacturer's instructions. Dissolve agonist compound in DMSO at the desired stock concentration, and prepare 3-fold serial dilutions of agonist compound in Cell Assay

Buffer. The concentration of each dilution should be prepared at 4× of the final screening concentration (i.e. 15 μL compound +45 μL Cell Assay Buffer/cAMP Antibody Reagent). For each dilution, the final concentration of solvent should remain constant. Transfer 15 μL diluted compound the assay plate and incubate the plate for 30 minutes at 37° C. Following agonist incubation, add 60 μL of working cAMP detection reagents/cAMP Solution mixture (cAMP Lysis Buffer, Substrate Reagent 1, cAMP Solution D) to the appropriate wells. Incubate for 1 hour at room temperature (23° C.), protected from light. Add 60 μl of cAMP Solution A to the appropriate wells. Incubate for 3 hours at room temperature (23° C.), protected from light. Read samples on Envision standard luminescence plate reader. Calculate of average EC₅₀ after logarithm transformation.

To assess the FXR agonistic potency of the example compounds as well as for reference compound, potency ranges were determined in the Human FXR (NR1H4) Assay as listed below in Table 2. The efficacy was normalized to CDCA set as 100%. (A=0.1 μM<EC50<1.0 μM; B=1.0 μM<EC50<10 μM; C=EC50>10 μM)

TABLE 2 Example EC50 (μM) Efficacy (%) CDCA C 100 6-ECDCA A 239 1 B 128 2 A 421 3 B 300 4 B 223 5 B 247 6 A 240 7 B 246 8 A 234 9 A 222 10 A 304 11 B 239 12 A 369 13 B 168 14 A 265 15 A 362 16 A 378 17 A 314 18 B 244 19 B 172 20 B 87 CDCA = chenodoxycholic acid; 6-ECDCA = 6-α-ethylchenodoxycholic acid.

While this invention has been particularly shown and described with references to preferred embodiments thereof, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the scope of the invention encompassed by the appended claims. 

1. A compound represented by Formula I or a pharmaceutically acceptable salt or ester thereof:

wherein: ring A is a five membered heteroaromatic ring comprising three heteroatoms independently selected from nitrogen, oxygen and sulfur, and is optionally further substituted; R₁ at each occurrence is independently selected from the group consisting of: 1) Hydrogen; 2) Substituted or unsubstituted —C₁-C₈ alkyl; 3) Substituted or unsubstituted —C₂-C₈ alkenyl; 4) Substituted or unsubstituted —C₂-C₈ alkynyl; 5) Substituted or unsubstituted aryl; and 6) Substituted or unsubstituted arylalkyl; m is selected from 0, 1, 2 and 3; R₂ is hydrogen, hydroxyl, —OSO₃H, —OSO₃ ⁻, —OAc, —OPO₃H₂ or —OPO₃ ²⁻; R₃ is hydrogen, halogen, CN, N₃, hydroxyl, —OSO₃H, —OSO₃ ⁻, —OAc, —OPOP₃H₂, —OPO₃ ²⁻, —SR₁ or —NHR₁, alternatively, R₂ and R₃ are taken together with the carbon atoms to which they are attached to form —CH═CH— or a cycloalkyl ring or heterocycloalkyl ring; R₄ and R₅ are each independently selected from hydrogen and hydroxyl protecting group; R₆ is selected from the group consisting of: 1) Hydrogen; 2) Halogen; 3) Substituted or unsubstituted —C₁-C₈ alkyl; 4) Substituted or unsubstituted —C₂-C₈ alkenyl; 5) Substituted or unsubstituted —C₂-C₈ alkynyl; and 6) Substituted or unsubstituted —C₃-C₈ cycloalkyl; and R₇ is selected from the group consisting of: 1) Hydrogen; 2) Substituted or unsubstituted —C₁-C₈ alkyl; 3) Substituted or unsubstituted —C₂-C₈ alkenyl; 4) Substituted or unsubstituted —C₂-C₈ alkynyl; 5) Substituted or unsubstituted —C₃-C₈ cycloalkyl; 6) Substituted or unsubstituted —C₃-C₈ heterocycloalkyl; 7) Substituted or unsubstituted aryl; 8) Substituted or unsubstituted heteroaryl; 9) Substituted or unsubstituted arylalkyl; 10) —NR₈R₉; 11) —N(R₁₀)C(O)R₁₁; 12) —N(R₁₀)C(O)OR₁₁; 13) —N(R₁₀)C(O)NR₈R₉; 14) —N(R₁₀)SO₂NR₈R₉; 15) —N(R₁₀)SO₂R₁₁; 16) —N(R₁₀)C(O)N(R₈)SO₂R₁₁; and 17) —N(R₁₀)C(O)N(R₈)SO₂NR₈R₉ R₈, R₉, R₁₀, and R₁₁ are each independently selected from hydrogen, substituted or unsubstituted —C₁-C₈ alkyl, substituted or unsubstituted —C₂-C₈ alkenyl, substituted or unsubstituted —C₂-C₈ alkynyl, substituted or unsubstituted —C₃-C₈ cycloalkyl, substituted or unsubstituted —C₃-C₈ heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted benzyl; alternatively, R₈ and R₉ or R₈ and R₁₀ are taken together with the atoms to which they are attached to form an optionally substituted heteroaryl ring.
 2. The compound of claim 1, wherein A-R₇ is selected from groups (I-1) to (I-11):

wherein R_(a) is selected from the group consisting of: 1) Hydrogen; 2) Substituted or unsubstituted —C₁-C₈ alkyl; 3) Substituted or un substituted —C₂-C₈ alkenyl; 4) Substituted or un substituted —C₂-C₈ alkynyl; 5) Substituted or unsubstituted —C₃-C₈ cycloalkyl; 6) —C(O)R₁₁; 7) —C(O)OR₁₁; 8) —C(O)NR₈R₉; 9) —SO₂R₁₁; and 10) —C(O)N(R₈)OS₂R₁₁; and R_(b) is is selected from the group consisting of: 1) Hydrogen; 2) Halogen; 3) CF₃; 4) CN; 5) Substituted or unsubstituted —C₁-C₈ alkyl; 6) Substituted or un substituted —C₂-C₈ alkenyl; 7) Substituted or un substituted —C₂-C₈ alkynyl; 8) Substituted or unsubstituted —C₃-C₈ cycloalkyl; 9) Substituted or unsubstituted —C₃-C₈ heterocycloalkyl; 10) Substituted or unsubstituted aryl; 11) Substituted or unsubstituted heteroaryl; 12) Substituted or unsubstituted arylalkyl; 13) —C(O)R₁₁; 14) —C(O)OR₁₁; 15) —C(O)NR₈R₉; and 16) —C(O)N(R₈)SO₂R₁₁.
 3. The compound of claim 1, represented by Formula II or a pharmaceutically acceptable salt or ester thereof:

wherein ring A, R₁, R₂, R₃, R₆, R₇ and m are as defined in claim
 1. 4. The compound of claim 1, represented by Formula III or a pharmaceutically acceptable salt or ester thereof:

wherein ring A, R₁, R₂, R₆, R₇ and m are as defined in claim
 1. 5. The compound of claim 1, represented by formula (III-1˜III-12), or a pharmaceutically acceptable salt or ester thereof,

wherein R₆, R₇ and m are as defined in claim
 1. 6. The compound of claim 1, represented by Formula IV or a pharmaceutically acceptable salt or ester thereof:

wherein R₇ and m are as defined in claim
 1. 7. The compound according to claim 6 selected from compounds of Formula IV wherein R₇ and m are delineated for each compound in Table 1: TABLE 1 compound m R₇ 1 0 H 2 0

3 0

4 0

5 0

6 0 —NH₂ 7 0 —NMe₂ 8 0

9 0

10 0

11 0

12 0

13 0

14 0

15 0

16 0

17 0

18 0

19 0

20 0

21 0

22 0

23 0

24 0

25 0

26 0

27 0

28 0

29 0

30 1 H 31 1

32 1

33 1

34 1

35 1 —NH₂ 36 1 —NMe₂ 37 1

38 1

39 1

40 1

41 1

42 1

43 1

44 1

45 1

46 1

47 1

48 1

49 1

50 1

51 1

52 1

53 1

54 1

55 1

56 1

57 1

58 1

59 2 H 60 2

61 2

62 2

63 2

64 2 —NH₂ 65 2 —NMe₂ 66 2

67 2

68 2

69 2

70 2

71 2

72 2

73 2

74 2

75 2

76 2

77 2

78 2

79 2

80 2

81 2

82 2

83 2

84 2

85 2

86 2

87 2

or a pharmaceutically acceptable salt or ester thereof.
 8. The compound of claim 1, represented by Formula V, or a pharmaceutically acceptable salt or ester thereof:

wherein R₇ and m are as defined in claim
 1. 9. The compound according to claim 8, selected from compounds of Formula V wherein R₇ and m are delineated for each compound in Table 2: TABLE 2 compound m R₇  88 0 H  89 0

 90 0

 91 0

 92 0

 93 0 —NH₂  94 0 —NMe₂  95 0

 96 0

 97 0

 98 0

 99 0

100 0

101 0

102 0

103 0

104 0

105 0

106 0

107 0

108 0

109 0

110 0

111 0

112 0

113 0

114 0

115 0

116 0

117 1 H 118 1

119 1

120 1

121 1

122 1 —NH₂ 123 1 —NMe₂ 124 1

125 1

126 1

127 1

128 1

129 1

130 1

131 1

132 1

133 1

134 1

135 1

136 1

137 1

138 1

139 1

140 1

141 1

142 1

143 1

144 1

145 1

146 2 H 147 2

148 2

149 2

150 2

151 2 —NH₂ 152 2 —NMe₂ 153 2

154 2

155 2

156 2

157 2

158 2

159 2

160 2

161 2

162 2

163 2

164 2

165 2

166 2

167 2

168 2

169 2

170 2

171 2

172 2

173 2

174 2

or a pharmaceutically acceptable salt or ester thereof.
 10. The compound of claim 1, represented by Formula VI or a pharmaceutically acceptable salt or ester thereof:

wherein R₇ and m are as defined in claim
 1. 11. The compound of claim 10, selected from compounds of Formula VI wherein R₇ and m are delineated for each compound in Table 3: TABLE 3 compound m R₇ 175 0 H 176 0

177 0

178 0

179 0

180 0 —NH₂ 181 0 —NMe₂ 182 0

183 0

184 0

185 0

186 0

187 0

188 0

189 0

190 0

191 0

192 0

193 0

194 0

195 0

196 0

197 0

198 0

199 0

200 0

201 0

202 0

203 0

204 1 H 205 1

206 1

207 1

208 1

209 1 —NH₂ 210 1 —NMe₂ 211 1

212 1

213 1

214 1

215 1

216 1

217 1

218 1

219 1

220 1

221 1

222 1

223 1

224 1

225 1

226 1

227 1

228 1

229 1

230 1

231 1

232 1

233 2 H 234 2

235 2

236 2

237 2

238 2 —NH₂ 239 2 —NMe₂ 240 2

241 2

242 2

243 2

244 2

245 2

246 2

247 2

248 2

249 2

250 2

251 2

252 2

253 2

254 2

255 2

256 2

257 2

258 2

259 2

260 2

261 2

or a pharmaceutically acceptable salt or ester thereof.
 12. The compound of claim 1, represented by Formula VII or a pharmaceutically acceptable salt or ester thereof:

wherein R₇ and m are previously defined in claim
 1. 13. The compound of claim 12, selected from compounds of Formula VII wherein R₇ and m are delineated for each compound in Table 4: TABLE 4 compound m R₇ 262 0 H 263 0

264 0

265 0

266 0

267 0 —NH₂ 268 0 —NMe₂ 269 0

270 0

271 0

272 0

273 0

274 0

275 0

276 0

277 0

278 0

279 0

280 0

281 0

282 0

283 0

284 0

285 0

286 0

287 0

288 0

289 0

290 0

291 1 H 292 1

293 1

294 1

295 1

296 1 —NH₂ 297 1 —NMe₂ 298 1

299 1

300 1

301 1

302 1

303 1

304 1

305 1

306 1

307 1

308 1

309 1

310 1

311 1

312 1

313 1

314 1

315 1

316 1

317 1

318 1

319 1

320 2 H 321 2

322 2

323 2

324 2

325 2 —NH₂ 326 2 —NMe₂ 327 2

328 2

329 2

330 2

331 2

332 2

333 2

334 2

335 2

336 2

337 2

338 2

339 2

340 2

341 2

342 2

343 2

344 2

345 2

346 2

347 2

348 2

or a pharmaceutically acceptable salt or ester thereof.
 14. A method for preventing or treating an FXR-mediated disease or condition in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound according to claim
 1. 15. The method according to claim 14, wherein the FXR-mediated disease or condition is selected from the group consisting of chronic liver disease, gastrointestinal disease, renal disease, cardiovascular disease, and metabolic disease.
 16. The method according to claim 15, wherein the chronic liver disease is selected from the group consisting of primary biliary cirrhosis (PBC), cerebrotendinous xanthomatosis (CTX), primary sclerosing cholangitis (PSC), drug induced cholestasis, intrahepatic cholestasis of pregnancy, parenteral nutrition associated cholestasis (PNAC), bacterial overgrowth or sepsis associated cholestasis, autoimmune hepatitis, chronic viral hepatitis, alcoholic liver disease, nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), liver transplant associated graft versus host disease, living donor transplant liver regeneration, congenital hepatic fibrosis, choledocholithiasis, granulomatous liver disease, intra- or extrahepatic malignancy, Sjogren's syndrome, Sarcoidosis, Wilson's disease, Gaucher's disease, hemochromatosis, and alpha 1-antitrypsin deficiency.
 17. The method according to claim 15, wherein the renal disease is selected from the group consisting of diabetic nephropathy, focal segmental glomerulosclerosis (FSGS), hypertensive nephrosclerosis, chronic glomerulonephritis, chronic transplant glomerulopathy, chronic interstitial nephritis, and polycystic kidney disease.
 18. The method according to claim 15, wherein the cardiovascular disease is selected from the group consisting of atherosclerosis, arteriosclerosis, dyslipidemia, hypercholesterolemia, and hypertriglyceridemia.
 19. The method according to claim 15, wherein the metabolic disease is selected from the group consisting of insulin resistance, Type I, diabetes, Type II diabetes, and obesity.
 20. A method for preventing or treating a TGR5-mediated disease or condition in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound according to claim
 1. 21. A pharmaceutical composition comprising a compound according to claim 1 and a pharmaceutically acceptable carrier.
 22. (canceled) 